Compositions, kits, and methods for anti-microbial serology assays using anti-human immunoglobulin antibody

Abstract

Reagents, kits, and microfluidics devices are disclosed for detecting the presence and/or concentration of antibodies directed to microorganisms in human biological samples. Also disclosed are methods of production and use of the reagents, kits, and microfluidics devices. Anti-human immunoglobulin antibodies are utilized as a reagent in a bridging immunoassay for detection of the microorganisms.

1 . A kit for performing a serology assay that detects the presence and/or concentration of antibodies to a microorganism in a human biological sample utilizing a chemiluminescent detection system, the kit comprising:
 (a) a composition comprising a singlet oxygen-activatable chemiluminescent compound having an antigen of the microorganism directly or indirectly bound thereto; and   (b) a composition comprising a sensitizer capable of generating singlet oxygen in its excited state and at least one anti-human immunoglobulin antibody directly or indirectly bound to the sensitizer.   
     
     
         2 . The kit of  claim 1 , wherein the microorganism is a virus. 
     
     
         3 . The kit of  claim 2 , wherein the virus is severe acute respiratory syndrome coronavirus 2 (SARS-COV-2). 
     
     
         4 . The kit of  claim 1 , wherein the at least one anti-human immunoglobulin antibody specifically binds to human IgA, human IgG, and human IgM. 
     
     
         5 . A kit for performing a serology assay that detects the presence and/or concentration of antibodies to a microorganism in a human biological sample utilizing a chemiluminescent detection system, the kit comprising:
 (a) a composition comprising a singlet oxygen-activatable chemiluminescent compound having an antigen of the microorganism directly or indirectly bound thereto;   (b) a composition comprising a sensitizer capable of generating singlet oxygen in its excited state and a biotin-specific binding partner directly or indirectly bound thereto; and   (c) at least one anti-human immunoglobulin antibody, wherein the antibody is biotinylated.   
     
     
         6 . The kit of  claim 5 , wherein the microorganism is a virus. 
     
     
         7 . The kit of  claim 6 , wherein the virus is severe acute respiratory syndrome coronavirus 2 (SARS-COV-2). 
     
     
         8 . The kit of  claim 5 , wherein the at least one anti-human immunoglobulin antibody specifically binds to human IgA, human IgG, and human IgM. 
     
     
         9 . A microfluidics device for detecting the presence and/or concentration of antibodies to a microorganism in a human biological sample, the microfluidics device comprising:
 (i) an inlet channel through which the human biological sample is applied;   (ii) at least a first compartment capable of being in fluidic communication with the inlet channel and containing:   (a) a composition comprising a singlet oxygen-activatable chemiluminescent compound having an antigen of the microorganism directly or indirectly bound thereto; and   (b) at least one composition comprising a sensitizer capable of generating singlet oxygen in its excited state, and at least one anti-human immunoglobulin antibody capable of being directly or indirectly bound to the sensitizer.   
     
     
         10 . The microfluidics device of  claim 9 , wherein the microorganism is a virus. 
     
     
         11 . The microfluidics device of  claim 10 , wherein the virus is severe acute respiratory syndrome coronavirus 2 (SARS-COV-2). 
     
     
         12 . The microfluidics device of  claim 9 , wherein the at least one anti-human immunoglobulin antibody of (b) specifically binds to human IgA, human IgG, and human IgM. 
     
     
         13 . The microfluidics device of  claim 9 , wherein (b) is further defined as:
 a composition comprising a sensitizer capable of generating singlet oxygen in its excited state and a biotin-specific binding partner directly or indirectly bound thereto; and   at least one anti-human immunoglobulin antibody, wherein the antibody is biotinylated.   
     
     
         14 . The microfluidics device of  claim 9 , wherein (a) and (b) are present in the same compartment. 
     
     
         15 . The microfluidics device of  claim 9 , wherein (a) and (b) are in two different compartments. 
     
     
         16 . A method for detecting the presence and/or concentration of antibodies to a microorganism in a human biological sample, the method comprising the steps of:
 (1) combining, either simultaneously or wholly or partially sequentially:   (a) the human biological sample suspected of containing antibodies to the microorganism;   (b) a composition comprising a singlet oxygen-activatable chemiluminescent compound having at least one antigen of the microorganism directly or indirectly bound thereto; and   (c) a composition comprising a sensitizer capable of generating singlet oxygen in its excited state and at least one anti-human immunoglobulin antibody directly or indirectly bound to the sensitizer;   (2) allowing the binding of (b) and (c) to antibodies to the microorganism present in (a), wherein the binding of (b) and (c) to the antibodies to the microorganism results in the formation of a complex in which the sensitizer is brought into close proximity to the chemiluminescent compound;   (3) activating the sensitizer to generate singlet oxygen, wherein activation of the sensitizers present in the complex causes the activation of the chemiluminescent compound present in each complex; and   (4) determining the amount of chemiluminescence generated by the activated chemiluminescent compound in the complex to determine the presence and/or concentration of antibodies to the microorganism present in the sample.   
     
     
         17 . The method of  claim 16 , wherein the microorganism is a virus. 
     
     
         18 . The method of  claim 17 , wherein the virus is severe acute respiratory syndrome coronavirus 2 (SARS-COV-2). 
     
     
         19 . The method of  claim 16 , wherein the at least one anti-human immunoglobulin antibody of (c) specifically binds to human IgA, human IgG, and human IgM. 
     
     
         20 . The method of  claim 16 , wherein the human biological sample is selected from the group consisting of whole blood or any portion thereof, urine, saliva, sputum, cerebrospinal fluid, skin, intestinal fluid, intraperitoneal fluid, cystic fluid, sweat, interstitial fluid, extracellular fluid, tears, mucus, bladder wash, semen, fecal, pleural fluid, nasopharyngeal fluid, and combinations thereof. 
     
     
         21 . A method for detecting the presence and/or concentration of antibodies to a microorganism in a human biological sample, the method comprising the steps of:
 (1) combining, either simultaneously or wholly or partially sequentially:   (a) the human biological sample suspected of containing antibodies to the microorganism;   (b) a composition comprising a singlet oxygen-activatable chemiluminescent compound having at least one antigen of the microorganism directly or indirectly bound thereto;   (c) a composition comprising a sensitizer capable of generating singlet oxygen in its excited state and a biotin-specific binding partner directly or indirectly bound thereto; and   (d) at least one anti-human immunoglobulin antibody, wherein the antibody is biotinylated;   (2) allowing the binding of (c) to (d) to form (c)/(d) and the binding of (b) and (c)/(d) to antibodies to the microorganism present in (a), wherein the binding of (b) and (c)/(d) to the antibodies to the microorganism results in the formation of a complex in which the sensitizer is brought into close proximity to the chemiluminescent compound;   (3) activating the sensitizer to generate singlet oxygen, wherein activation of the sensitizers present in the complex causes the activation of the chemiluminescent compound present in each complex; and   (4) determining the amount of chemiluminescence generated by the activated chemiluminescent compound in the complex to determine the presence and/or concentration of antibodies to the microorganism present in the sample.   
     
     
         22 . The method of  claim 21 , wherein the microorganism is a virus. 
     
     
         23 . The method of  claim 22 , wherein the virus is severe acute respiratory syndrome coronavirus 2 (SARS-COV-2). 
     
     
         24 . The method of  claim 21 , wherein the at least one anti-human immunoglobulin antibody of (c) specifically binds to human IgA, human IgG, and human IgM. 
     
     
         25 . The method of  claim 21 , wherein the human biological sample is selected from the group consisting of whole blood or any portion thereof, urine, saliva, sputum, cerebrospinal fluid, skin, intestinal fluid, intraperitoneal fluid, cystic fluid, sweat, interstitial fluid, extracellular fluid, tears, mucus, bladder wash, semen, fecal, pleural fluid, nasopharyngeal fluid, and combinations thereof. 
     
     
         26 .- 50 . (canceled)